摘要
目的观察沉默BC047440基因后对HepG2细胞侵袭能力的影响。方法实验分为3组,即HepG2细胞转染siRNA的沉默组,HepG2细胞转染无关质粒组及未转染的HepG2细胞组。利用前期构建的BC047440基因的小干扰RNA(siRNA)沉默BC047440基因,然后用MTT法检测细胞黏附率,细胞划痕法检测细胞迁移能力,Boyden小室法检测细胞侵袭性。结果沉默组的细胞黏附率在20 min和40 min时与HepG2细胞组和无关质粒组比较,差异无统计学意义(P>0.05),但在60 min时沉默组显著低于后2组(P<0.05);划痕实验显示划痕48h后,无关质粒组和HepG2细胞组划痕区已基本长满,而沉默组划痕依然明显;接种16 h后无关质粒组和HepG2细胞组侵袭到Boyden小室下室面的细胞数明显多于沉默(P<0.05)。而无关质粒组与HepG2细胞组间差异均无显著性(P>0.05)。结论沉默BC047440基因后可明显抑制HepG2细胞其在细胞外基质上的黏附能力、运动能力和体外侵袭能力,提示BC047440基因可增加肝癌的转移侵袭性。
Objective To investigate the change of the invasive and metastasis ability of HepG2 cells by silencing BC047440 genes. Methods The experiment were divided in 3 groups： in terfering group, HepG2/HK group and HepG2 group. Small interfering RNAs for BC047440 gene were initially designed and silenced to BC047440 genes, then the cell adhesion was measured with MTT assay, the cell migration was surveyed by scratch method, and the cell invasion in vitro was determined with Boyden chamber. Results There was no significant difference of the adhesion rate in the interfering group compared with HepG2/HK group and HepG2 group at 20 min and 40 min after seeding, while there was significant difference at 60min （P 〈 0.05 ）. The scratch wound had almost been filled with HepG2/HK group and HepG2 group at 48h after scratching, while it was still apparent in the interfering group. The number of ceils that penetrated polycarbonate coated with matrigel when incubation for 16h in interfering group was much less than HepG2/ HK group and HepG2 group （ P 〈 0.05 ）. Conclusions The silencing BC047440 genes could significantly inhibit the ability of adhesion to extracellular matrix, the ability of migration to scratch wound, the ability of invasion of HepG2 cells in vitro, and these results suggest that BC047440 could regulate the invasive and metastasis ability of HepG2 cells.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2009年第1期49-52,共4页
China Journal of General Surgery
基金
国家自然科学基金资助项目(30570843)
